Search results for: bacterial consortium
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1226

Search results for: bacterial consortium

236 Impact of a Structured Antimicrobial Stewardship Program in a North-East Italian Hospital

Authors: Antonio Marco Miotti, Antonella Ruffatto, Giampaola Basso, Antonio Madia, Giulia Zavatta, Emanuela Salvatico, Emanuela Zilli

Abstract:

A National Action Plan to fight antimicrobial resistance was launched in Italy in 2017. In order to reduce inappropriate exposure to antibiotics and infections from multi-drug resistant bacteria, it is essential to set up a structured system of surveillance and monitoring of the implementation of National Action Plan standards, including antimicrobial consumption, with a special focus on quinolones, third generation cephalosporins and carbapenems. A quantitative estimate of antibiotic consumption (defined daily dose - DDD - consumption per 100 days of hospitalization) has been provided by the Pharmaceutical Service to the Hospital of Cittadella, ULSS 6 Euganea – Health Trust (District of Padua) for the years 2019 (before the pandemic), 2020 and 2021 for all classes of antibiotics. Multidisciplinary meetings have been organized monthly by the local Antimicrobial Stewardship Group. Between 2019 and 2021, an increase in the consumption of carbapenems in the Intensive Care Unit (from 12.2 to 18.2 DDD, + 49.2%) and a decrease in Medical wards (from 5.3 to 2.6 DDD, - 50.9%) was reported; a decrease in the consumption of quinolones in Intensive Care Unit (from 17.2 to 10.8 DDD, - 37.2%), Medical wards (from 10.5 to 6.6 DDD, - 37.1%) and Surgical wards (from 10.2 to 9.3 DDD, - 8.8%) was highlighted; an increase in the consumption of third generation cephalosporins in Medical wards (from 18.1 to 22.6 DDD, + 24,1%) was reported. Finally, after an increase in the consumption of macrolides between 2020 and 2019, in 2021, a decrease was reported in the Intensive Care Unit (DDD: 8.0 in 2019, 18.0 in 2020, 6.4 in 2021) and Medical wards (DDD: 9.0 in 2019, 13.7 in 2020, 10.9 in 2021). Constant monitoring of antimicrobial consumption and timely identifying of warning situations that may need a specific intervention are the cornerstone of Antimicrobial Stewardship programs, together with analysing data on bacterial resistance rates and infections from multi-drug resistant bacteria.

Keywords: carbapenems, quinolones, antimicrobial, stewardship

Procedia PDF Downloads 158
235 Satureja bachtiarica Bunge Induce Apoptosis via Mitochondrial Intrinsic Pathway and G1 Cell Cycle Arrest

Authors: Hamed Karimian, Noraziah Nordin, Mohamad Ibrahim Noordin, Syam Mohan, Mahboubeh Razavi, Najihah Mohd Hashim, Happipah Mohd Ali

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Satureja bachtiarica Bunge is a perennial medicinal plant belonging to the Lamiaceae family and endemic species in Iran. Satureja bachtiarica Bunge with the local name of Marzeh koohi is edible vegetable use as flavoring agent, anti-bacterial and to relieve cough and indigestion. In this study, the anti-cancer effect of Satureja bachtiarica Bunge on the MDA-MB-231 cell line as an Breast cancer cell model has been analyzed for the first time. Satureja bachtiarica Bunge was extracted using different solvents in the order of increasing polarity. Cytotoxicity activity of hexane extract of Satureja bachtiarica Bunge (SBHE) was observed using MTT assay. Acridine orange/Propidium iodide staining was used to detect early apoptosis; Annexin-V-FITC assay was carried out to observe the detection of cell-surface Phosphatidylserine (PS), with Annexin-Vserving as a marker for apoptotic cells. Caspase 3/7, 8 and-9 assays showed significantly activation of caspase-9 where lead intrinsic mitochondrial pathway. Bcl-2/Bax expressions and cell cycle arrest were also investigated. SBHE had exhibited significantly higher cytotoxicity against MDA-MB-231 Cell line compare to other cell lines. A significant increase in chromatin condensation in the cell nucleus was observed by fluorescence analysis. Treatment of MDA-MB-231 cells with SBHE encouraged apoptosis, by down-regulating Bcl-2 and up-regulating Bax, which lead the activation of caspase 9. Moreover, SBHE treatment significantly arrested MDA-MB-231 cells in the G1 phase. Together, the results presented in this study demonstrated that SBHE inhibited the proliferation of MDA-MB-231 cells, leading cell cycle arrest and programmed cell death, which was confirmed to be through the mitochondrial pathway.

Keywords: Satureja bachtiarica Bunge, MDA-MB-231, apoptosis, annexin-V, cell cycle

Procedia PDF Downloads 337
234 Selective Recovery and Molecular Identification of Laccase-Producing Bacteria from Selected Terrestrial and Aquatic Milieu in the Eastern Cape, South Africa: Toward the Production of Environmentally Relevant Biocatalysts

Authors: John Onolame Unuofin, Uchechukuw U. Nwodo, Anthony I. Okoh

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Laccase is constantly gaining status as important biocatalyst in biotechnology. The illimitable potential of its industrial applications and the corresponding aggressive need for phenomenal volumes of extracellularly secreted laccases have called for its interminable production from sources which are able to meet this demand within a relatively short period of time, preferably bacteria. In response to this call, this study was designed to source for laccase-producing bacteria from different environmental matrices. Three sampling environments were chosen such as wastewater treatment plants, University of Fort Hare vicinity and the Hogback woodland, all within the Eastern Cape, South Africa. Samples such as effluents, sediments, leaf litters, degrading wood and rock scrapings were selectively enriched with some model aromatic compounds and were further screened qualitatively and quantitatively on five phenolic substrates ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), Guaiacol, 1-Naphthol, Potassium Ferric Cyanide and Syringaldazine). Basis for selection was their ability to elicit a colour change on at least three of the above mentioned agar based assay substrates. The choice isolates were further identified based on 16S rRNA molecular identification techniques. 33 isolates were screened out of the 40 representative distinct colonies during the qualitative plate screens, while quantitative screens selected out 11 bacterial isolates. They were, based on molecular identification, desginated as members of the genera Pseudomonas, Stenotrophomonas and Citrobacter of the gammaproteobacteria and Bordetalla and Achromobacter of the betaproteobacteria respectively. We therefore conclude based on our outcomes that we may have isolated efficient laccase-producing bacteria, which might be of beneficial significance in catalysis and biotechnology.

Keywords: beta proteobacteria, catalysis, gammaproteobacteria, laccase

Procedia PDF Downloads 174
233 Effect of Media Osmolarity on Vi Biosynthesis on Salmonella enterica serovar Typhi Strain C6524 Cultured on Batch System

Authors: Dwi Arisandi Wijaya, Ernawati Arifin Giri-Rachman, Neni Nurainy

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Typhoid fever disease can be prevented by using a polysaccharide-based vaccine Vi which is a virulence factor of S.typhi. To produce high yield Vi polysaccharide from bacteria, it is important to know the biosynthesis of Vi polysaccharide and the regulators involved. In the In vivo condition, S. typhi faces different osmolarity, and the bacterial two-component system OmpR-EnvZ, regulate by up and down Capsular Vi polysaccharide biosynthesis. A high yielded Vi Polysaccharide strain, S. typhi strain C6524 used to study the effect of media osmolarity on Vi polysaccharide biosynthesis and the osmoregulation pattern of S. typhi strain C6524. The methods were performed by grown S. typhi strain C6524 grown on medium with 50 mM, 100 mM, and 150 mM osmolarity with the batch system. Vi polysaccharide concentration was measured by ELISA method. For further investigation of the osmoregulation pattern of strain C6524, the osmoregulator gene, OmpR, has been isolated and sequenced using the specific primer of the OmpR gene. Nucleotide sequence analysis is done with BLAST and Lallign. Amino Acid sequence analysis is done with Prosite and Multiple Sequence Alignment. The results of cultivation showed the average content of polysaccharide Vi for 50 mM, 100 mM, and 150 mM osmolarities 11.49 μg/mL, 12.06 μg/mL, and 14.53 μg/mL respectively. Analysis using Anova stated that the osmolarity treatment of 150 mM significantly affects Vi content. Analysis of nucleotide sequences shows 100% identity between S. typhi strain C6524 and Ty2. Analysis of amino acid sequences shows that the OmpR response regulator protein of the C6524 strain also has a α4-β5-α5 motif which is important for the regulatory activation system when phosphorylation occurs by domain kinase. This indicates that the regulator osmolarity response of S. typhi strain C6524 has no difference with the response regulator owned by S. typhi strain Ty2. A high Vi response rate in the 150 mM osmolarity treatment requires further research for RcsB-RcsC, another two-component system involved in Vi Biosynthesis.

Keywords: osmoregulator, OmpR, Salmonella, Vi polysaccharide

Procedia PDF Downloads 198
232 Microbial Quality Assessment of Indian White Shrimp, Penaeus Indicus from Southwest Bangladesh

Authors: Saima Sharif Nilla, Mahmudur Rahman Khan, Anisur Rahman Khan, Ghulam Mustafa1

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The microbial quality of Indian white shrimp (Peneaus indicus) from Bagerhat, Khulna and Satkhira of southwest Bangladesh was assessed where the parameters varied with different sources and the quality was found to be poor for Satkhira shrimp samples. Shrimp samples in fresh condition were collected to perform the microbial assessment and 10 pathogenic isolates for antibiotic sensitivity test to 12 antibiotics. The results show that total bacterial count of all the samples were beyond the acceptable limit 105 cfu/g. In case of total coliform and E. coli density, no substantial difference (p<0.5) was found between the different shrimp samples from different districts and also high quantity of TC exceeding the limit (>102 cfu/g) proves the poor quality of shrimp. The FC abundance found in shrimps of Bagerhat and Satkhira was similar and significantly higher (p<0.5) than that of Khulna samples. No significant difference (p<0.5) was found among the high density of Salmonella-Shigella, Vibrio spp., and Staphylococcus spp. of the shrimp samples from the source places. In case of antibiotic sensitivity patterns, all of them were resistant to ampicillin, Penicillin and sensitive to kanamycin. Most of the isolates were frequently sensitive to ciprofloxacin and streptomycin in the sensitivity test. In case of nutritional composition, no significant difference (t-test, p<0.05) was found among protein, lipid, moisture and ash contents of shrimp samples. The findings prove that shrimp under this study was more or less contaminated and samples from Satkhira were highly privileged with food borne pathogens which confirmed the unhygienic condition of the shrimp farms as well as the presence of antibiotic resistance bacteria in shrimp fish supposed to threat food safety and deteriorate the export quality.

Keywords: food borne pathogens, satkhira, penaeus indicus, antibiotic sensitivity, southwest Bangladesh, food safety

Procedia PDF Downloads 708
231 Theory of Negative Trigger: The Contract between Oral Probiotics and Immune System

Authors: Cliff Shunsheng Han

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Identifying the direct allergy cause that can be easily mitigated is the foundation to stop the allergy epidemic that has been started in the seventies. It has confirmed that the personal and social hygiene practices are associated with the allergy prevalence. But direct causes have been found, and proposed translational measures have not been effective. This study, assisted by a particular case of allergies, has seen the direct cause of allergies, developed a valid test resulted in lasting relief for allergies, and constructed theory describing general relationship between microbiota and host immune system. Saliva samples were collected from a subject for three years during which time the person experienced yearlong allergy, seasonal allergy, and remission of allergy symptoms. Bacterial DNA was extracted and 16S rRNA genes were profiled with Illumina sequencing technology. The analyzing results indicate that the possible direct cause of allergy is the lacking probiotic bacteria in the oral cavity, such as genera Streptococcus and Veilonella, that can produce metabolites to pacify immune system. Targeted promotion of those bacteria with a compound designed for them, has led to lasting remissions of allergic rhinitis. During the development of the translational measure, the subject's oral biofilm was completely destructed by a moderate fever due to an unrelated respiratory infection. The incident not only facilitated the development of the heat based microbiota reseeding procedure but also indicated a possible natural switch that subsequently increases the efficacy of the immune system previously restrained by metabolites from microbiota. These results lead to the proposal of a Theory of Negative Trigger (TNT) to describe the relationship between oral probiotics and immune system, in which probiotics are the negative trigger that will release the power of immune system when removed by fever or modern lifestyles. This study could open doors leading to further understanding of how the immune system functions under the influence of microbiota as well as validate simple traditional practices for healthy living.

Keywords: oral microbiome, allergy, immune system, infection

Procedia PDF Downloads 131
230 Greening the Blue: Enzymatic Degradation of Commercially Important Biopolymer Dextran Using Dextranase from Bacillus Licheniformis KIBGE-IB25

Authors: Rashida Rahmat Zohra, Afsheen Aman, Shah Ali Ul Qader

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Commercially important biopolymer, dextran, is enzymatically degraded into lower molecular weight fractions of vast industrial potential. Various organisms are associated with dextranase production, among which fungal, yeast and bacterial origins are used for commercial production. Dextranases are used to remove contaminating dextran in sugar processing industry and also used in oral care products for efficient removal of dental plaque. Among the hydrolytic products of dextran, isomaltooligosaccharides have prebiotic effect in humans and reduces the cariogenic effect of sucrose in oral cavity. Dextran derivatives produced by hydrolysis of high molecular polymer are also conjugated with other chemical and metallic compounds for usage in pharmaceutical, fine chemical industry, cosmetics, and food industry. Owing to the vast application of dextran and dextranases, current study focused on purification and analysis of kinetic parameters of dextranase from a newly isolated strain of Bacillus licheniformis KIBGE-IB25. Dextranase was purified up to 35.75 folds with specific activity of 1405 U/mg and molecular weight of 158 kDa. Analysis of kinetic parameters revealed that dextranase performs optimum cleavage of low molecular weight dextran (5000 Da, 0.5%) at 35ºC in 15 min at pH 4.5 with a Km and Vmax of 0.3738 mg/ml and 182.0 µmol/min, respectively. Thermal stability profiling of dextranase showed that it retained 80% activity up to 6 hours at 30-35ºC and remains 90% active at pH 4.5. In short, the dextranase reported here performs rapid cleavage of substrate at mild operational conditions which makes it an ideal candidate for dextran removal in sugar processing industry and for commercial production of low molecular weight oligosaccharides.

Keywords: Bacillus licheniformis, dextranase, gel permeation chromatograpy, enzyme purification, enzyme kinetics

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229 Designing Function Knitted and Woven Upholstery Textile With SCOPY Film

Authors: Manar Y. Abd El-Aziz, Alyaa E. Morgham, Amira A. El-Fallal, Heba Tolla E. Abo El Naga

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Different textile materials are usually used in upholstery. However, upholstery parts may become unhealthy when dust accrues and bacteria raise on the surface, which negatively affects the user's health. Also, leather and artificial leather were used in upholstery but, leather has a high cost and artificial leather has a potential chemical risk for users. Researchers have advanced vegie leather made from bacterial cellulose a symbiotic culture of bacteria and yeast (SCOBY). SCOBY remains a gelatinous, cellulose biofilm discovered floating at the air-liquid interface of the container. But this leather still needs some enhancement for its mechanical properties. This study aimed to prepare SCOBY, produce bamboo rib knitted fabrics with two different stitch densities, and cotton woven fabric then laminate these fabrics with the prepared SCOBY film to enhance the mechanical properties of the SCOBY leather at the same time; add anti-microbial function to the prepared fabrics. Laboratory tests were conducted on the produced samples, including tests for function properties; anti-microbial, thermal conductivity and light transparency. Physical properties; thickness and mass per unit. Mechanical properties; elongation, tensile strength, young modulus, and peel force. The results showed that the type of the fabric affected significantly SCOBY properties. According to the test results, the bamboo knitted fabric with higher stitch density laminated with SCOBY was chosen for its tensile strength and elongation as the upholstery of a bed model with antimicrobial properties and comfortability in the headrest design. Also, the single layer of SCOBY was chosen regarding light transparency and lower thermal conductivity for the creation of a lighting unit built into the bed headboard.

Keywords: anti-microbial, bamboo, rib, SCOPY, upholstery

Procedia PDF Downloads 64
228 Antimicrobial, Antioxidant and Cytotoxic Activities of Cleoma viscosa Linn. Crude Extracts

Authors: Suttijit Sriwatcharakul

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The bioactivity studies from the weed ethanolic crude extracts from leaf, stem, pod and root of wild spider flower; Cleoma viscosa Linn. were analyzed for the growth inhibition of 6 bacterial species; Salmonella typhimurium TISTR 5562, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus TISTR 1466, Streptococcus epidermidis ATCC 1228, Escherichia coli DMST 4212 and Bacillus subtilis ATCC 6633 with initial concentration crude extract of 50 mg/ml. The agar well diffusion results found that the extracts inhibit only gram positive bacteria species; S. aureus, S. epidermidis and B. subtilis. The minimum inhibition concentration study with gram positive strains revealed that leaf crude extract give the best result of the lowest concentration compared with other plant parts to inhibit the growth of S. aureus, S. epidermidis and B. subtilis at 0.78, 0.39 and lower than 0.39 mg/ml, respectively. The determination of total phenolic compounds in the crude extracts exhibited the highest phenolic content was 10.41 mg GAE/g dry weight in leaf crude extract. Analyzed the efficacy of free radical scavenging by using DPPH radical scavenging assay with all crude extracts showed value of IC50 of leaf, stem, pod and root crude extracts were 8.32, 12.26, 21.62 and 35.99 mg/ml, respectively. Studied cytotoxicity of crude extracts on human breast adenocarcinoma cell line by MTT assay found that pod extract had the most cytotoxicity CC50 value, 32.41 µg/ml. Antioxidant activity and cytotoxicity of crude extracts exhibited that the more increase of extract concentration, the more activities indicated. According to the bioactivities results, the leaf crude extract of Cleoma viscosa Linn. is the most interesting plant part for further work to search the beneficial of this weed.

Keywords: antimicrobial, antioxidant activity, Cleoma viscosa Linn., cytotoxicity test, total phenolic compound

Procedia PDF Downloads 273
227 Functional Significance of Qatari Camels Milk: Antioxidant Content and Antimicrobial Activity of Protein Fractions

Authors: Tahra ElObeid, Omnya Ahmed, Reem Al-Sharshani, Doaa Dalloul, Jannat Alnattei

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Background: Camelus dormedarius camels are also called ‘the Arabian camels’ and are present in the desert area of North Africa and the Middle East. Recently, camel’s milk has a great attention globally because of their proteins and peptides that have been reported to be beneficial for the health and in the management of many diseases. Objectives: This study was designed to investigate the antioxidant, antimicrobial activity and to evaluate the total phenolic content of camel’s milk proteins in Qatar. Methods: Fresh two camel’s milk samples from Omani breed and called Muhajer (camel’s milk A and B) were collected on the 1st of the December. Both samples were from the same location Al- Shahaniyah, Doha, Qatar, but from different local private farms and feeding system. Camel’s milk A and B were defatted by centrifugation and their proteins were extracted by acid and thermal precipitation. The antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Total phenolic compound (TPC) was evaluated by Folin-Ciocalteu reagent (FCR). On the other hand, the antimicrobial activity against eight different type of pathogenic bacteria was evaluated by disc diffusion method and the zone of inhibition was measured. Results: The of the total phenolic content of whole milk in both camel’s milk A and B were significantly the highest among the protein extracts. The % of the DPPH radical inhibition of casein protein in both camel’s milk A and B were significantly the highest among the protein extracts. In this study, there were marked changes in the antibacterial activity in the different camel milk protein extracts. All extracts showed bacterial overgrowth. Conclusion: The antioxidant activity of the camel milk protein extracts correlated to their unique phenolic compounds and bioactive protein peptides. The antimicrobial activity was not detected perhaps due to the technique, the quality, or the extraction method. Overall, camel's milk exhibits a high antioxidant activity, which is responsible for many health benefits besides the nutritional values.

Keywords: camels milk, antioxidant content, antimicrobial activity, proteins, Qatar

Procedia PDF Downloads 215
226 Non-Candida Albicans Candida: Virulence Factors and Species Identification in India

Authors: Satender Saraswat, Dharmendra Prasad Singh, Rajesh Kumar Verma, Swati Sarswat

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Background and Purpose: The predominant cause of candidiasis was Candida albicans which has shifted towards non-Candida albicans Candida (NCAC) (Candida species other than the C. albicans). NCAC, earlier considered non-pathogenic or minimally virulent, are now considered a primary cause of morbidity and mortality in immunocompromised. With the NCAC spp. gaining weightage in the clinical cases, this study was conducted to determine the prevalence of NCAC spp. in different clinical specimens and to assess a few of their virulence factors. Material and Methods: Routine samples for bacterial culture and sensitivity, showing colony characteristics like Candida on Blood Agar and microscopic features resembling Candida spp. were processed further. Candida isolates were tested for chlamydospore formation, biochemical tests including sugar fermentation and sugar assimilation tests, and growth at 42oC, colony colour on HiCrome™ Candida Differential Agar, HiCandida Identification Kit and VITEK-2 Compact. Virulence factors like adherence to buccal epithelial cells (ABEC), biofilm formation, hemolytic activity, and production of coagulase enzyme were also tested. Results: Mean age of the patients was 38.46 with a male-female ratio of 1.36:1. 137 Candida isolates were recovered. 45.3% isolates were isolated from urine, 19.7% from vaginal swabs and 13.9% from oropharyngeal swabs. 55 (40.1%) isolates of C. albicans and 82 (59.9%) of NCAC spp. were identified, with C. tropicalis (23.4%) in NCAC. C. albicans (3; 50%) was the commonest species in cases of candidemia. Haemolysin production (85.5%) and ABEC (78.2%) were the major virulence factors in C. albicans. C. tropicalis (59.4%) and C. dubliniensis (50%) showed maximum ABEC. Biofilm forming capacity was higher in C. tropicalis (78.1%) than C. albicans (67%). Conclusion: This study suggests varied prevalence and virulence based on geographical locations, even within a subcontinent. It clearly demarcates the emergence of NCAC and their predominance in different body fluids. Identification of Candida to species level should become a routine in all the laboratories.

Keywords: ABEC, NCAC, non-Candida albicans Candida, Vitek-2TM compact

Procedia PDF Downloads 134
225 Silver Nanoparticles Synthesized in Plant Extract Against Acute Hepatopancreatic Necrosis of Shrimp: Estimated By Multiple Models

Authors: Luz del Carmen Rubí Félix Peña, Jose Adan Felix-Ortiz, Ely Sara Lopez-Alvarez, Wenceslao Valenzuela-Quiñonez

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On a global scale, Mexico is the sixth largest producer of farmed white shrimp (Penaeus vannamei). The activity suffered significant economic losses due to acute hepatopancreatic necrosis (AHPND) caused by a strain of Vibrio parahaemolyticus. For control, the first option is the application of antibiotics in food, causing changes in the environment and bacterial communities, which has produced greater virulence and resistance of pathogenic bacteria. An alternative treatment is silver nanoparticles (AgNPs) generated by green synthesis, which have shown an antibacterial capacity by destroying the cell membrane or denaturing the cell. However, the doses at which these are effective are still unknown. The aim is to calculate the minimum inhibitory concentration (MIC) using the Gompertz, Richard, and Logistic model of biosynthesized AgNPs against a strain of V. parahaemolyticus. Through the testing of different formulations of AgNPs synthesized from Euphorbia prostrate (Ep) extracts against V. parahaemolyticus causing AHPND in white shrimp. Aqueous and ethanol extracts were obtained, and the concentration of phenols and flavonoids was quantified. In the antibiograms, AgNPs were formulated in ethanol extracts of Ep (20 and 30%). The inhibition halo at well dilution test were 18±1.7 and 17.67±2.1 mm against V. parahaemolyticus. A broth microdilution was performed with the inhibitory agents (aqueous and ethanolic extracts and AgNPs) and 20 μL of the inoculum of V. parahaemolyticus. The MIC for AgNPs was 6.2-9.3 μg/mL and for ethanol extract of 49-73 mg/mL. The Akaike index (AIC) was used to choose the Gompertz model for ethanol extracts of Ep as the best data descriptor (AIC=204.8, 10%; 45.5, 20%, and 204.8, 30%). The Richards model was at AgNPs ethanol extract with AIC=-9.3 (10%), -17.5 (20 and 30%). The MIC calculated for EP extracts with the modified Gompertz model were 20 mg/mL (10% and 20% extract) and 40 mg/mL at 30%, while Richard was winner for AgNPs-synthesized it was 5 μg/mL (10% and 20%) and 8 μg/mL (30%). The solver tool Excel was used for the calculations of the models and inhibition curves against V.parahaemolyticus.

Keywords: green synthesis, euphorbia prostata, phenols, flavonoids, bactericide

Procedia PDF Downloads 106
224 Rapid and Cheap Test for Detection of Streptococcus pyogenes and Streptococcus pneumoniae with Antibiotic Resistance Identification

Authors: Marta Skwarecka, Patrycja Bloch, Rafal Walkusz, Oliwia Urbanowicz, Grzegorz Zielinski, Sabina Zoledowska, Dawid Nidzworski

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Upper respiratory tract infections are one of the most common reasons for visiting a general doctor. Streptococci are the most common bacterial etiological factors in these infections. There are many different types of Streptococci and infections vary in severity from mild throat infections to pneumonia. For example, S. pyogenes mainly contributes to acute pharyngitis, palatine tonsils and scarlet fever, whereas S. Streptococcus pneumoniae is responsible for several invasive diseases like sepsis, meningitis or pneumonia with high mortality and dangerous complications. There are only a few diagnostic tests designed for detection Streptococci from the infected throat of patients. However, they are mostly based on lateral flow techniques, and they are not used as a standard due to their low sensitivity. The diagnostic standard is to culture patients throat swab on semi selective media in order to multiply pure etiological agent of infection and subsequently to perform antibiogram, which takes several days from the patients visit in the clinic. Therefore, the aim of our studies is to develop and implement to the market a Point of Care device for the rapid identification of Streptococcus pyogenes and Streptococcus pneumoniae with simultaneous identification of antibiotic resistance genes. In the course of our research, we successfully selected genes for to-species identification of Streptococci and genes encoding antibiotic resistance proteins. We have developed a reaction to amplify these genes, which allows detecting the presence of S. pyogenes or S. pneumoniae followed by testing their resistance to erythromycin, chloramphenicol and tetracycline. What is more, the detection of β-lactamase-encoding genes that could protect Streptococci against antibiotics from the ampicillin group, which are widely used in the treatment of this type of infection is also developed. The test is carried out directly from the patients' swab, and the results are available after 20 to 30 minutes after sample subjection, which could be performed during the medical visit.

Keywords: antibiotic resistance, Streptococci, respiratory infections, diagnostic test

Procedia PDF Downloads 129
223 A Study of the Effects of Temperatures and Optimum pH on the Specific Methane Production of Perennial Ryegrass during Anaerobic Digestion Process under a Discontinuous Daily Feeding Condition

Authors: Uchenna Egwu, Paul Jonathan Sallis

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Perennial ryegrass is an abundant renewable lignocellulosic biofuel feedstock for biomethane production through anaerobic digestion (AD). In this study, six anaerobic continuously stirred tank reactors (CSTRs) were set up in three pairs. Each pair of the CSTRs was then used to study the effects of operating temperatures – psychrophilic, mesophilic, and thermophilic, and optimum pH on the specific methane production (SMP) of the ryegrass during AD under discontinuous daily feeding conditions. The reactors were fed at an organic loading rate (OLR) ranging from 1-1.5 kgVS.L⁻¹d⁻¹ and hydraulic residence time, HRT=20 days for 140 days. The pH of the digesters was maintained at the range of 6.8-7.2 using 1 M NH₄HCO₃ solution, but this was replaced with biomass ash-extracts from day 105-140. The results obtained showed that the mean SMP of ryegrass measured between HRT 3 and 4 were 318.4, 425.4 and 335 N L CH₄ kg⁻¹VS.d⁻¹ for the psychrophilic (25 ± 2°C), mesophilic (40 ± 1°C) and thermophilic (60 ± 1°C) temperatures respectively. It was also observed that the buffering ability of the reactors increased with operating temperature, probably due to an increase in the solubility of ammonium bicarbonate (NH₄HCO₃) with temperature. The reactors also achieved a mean VS destruction of 61.9, 68.5 and 63.5%, respectively, which signifies that the mesophilic reactors achieved the highest specific methane production (SMP), while the psychrophilic reactors achieved the lowest. None of the reactors attained steady-state condition due to the discontinuous daily feeding times, and therefore, such feeding practice may not be the most effective for maximum biogas production over long periods of time. The addition of NH₄HCO₃ as supplement provided a good buffering condition in these AD digesters, but the digesters failed in the long run due to inhibition from the accumulation of free ammonia, which later led to decrease in pH, acidification, and souring of the digesters. However, the addition of biomass ash extracts was shown to potentially revive failed AD reactors by providing an adequate buffering and essential trace nutrient supplements necessary for optimal bacterial growth.

Keywords: anaerobic digestion, discontinuous feeding, perennial ryegrass, specific methane production, supplements, temperature

Procedia PDF Downloads 127
222 Potential Application of Thyme (Thymus vulgaris L.) Essential Oil as Antibacterial Drug in Aromatherapy

Authors: Ferhat Mohamed Amine, Boukhatem Mohamed Nadjib, Chemat Farid

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The Lamiaceae family is widely spread in Algeria. Due to the application of Thymus species growing wild in Algeria as a culinary herb and in folk medicine, the purpose of the present work was to evaluate antimicrobial activities of their essential oils and relate them with their chemical composition, for further application in food and pharmaceutical industries as natural valuable products. The extraction of the Thymus vulgaris L. essential oil (TVEO) was obtained by steam distillation. Chemical composition of the TVEO was determined by Gas Chromatography. A total of thirteen compounds were identified. Carvacrol (83.8%) was the major component, followed by cymene (8.15%) and terpinene (4.96%). Antibacterial action of the TVEO against 23 clinically isolated bacterial strains was determined by using agar disc diffusion and vapour diffusion methods at different doses. By disc diffusion method, TVEO showed potent antimicrobial activity against gram-positive bacteria more than gram-negative strains and antibiotic discs. The Diameter of Inhibition Zone (DIZ) varied from 25 to 60 mm for S. aureus, B. subtilisand E. coli. However, the results obtained by both agar diffusion and vapour diffusion methods were different. Significantly higher antibacterial effect was observed in the vapour phase at lower doses. S. aureus and B. subtilis were the most susceptible strains to the oil vapour. Therefore, smaller doses of EO in the vapour phase can be inhibitory to pathogenic bacteria. There is growing evidence that TVEO in vapour phase are effective antiseptic systems and appears worthy to be considered for practical uses in the treatment of human infections oras air decontaminants in hospital. TVEO has considerable antibacterial activity deserving further investigation for clinical applications. Also whilst the mode of action remains mainly undetermined, this experimental approach will need to continue.

Keywords: antimicrobial drugs, carvacrol, disc diffusion, Thymus vulgaris, vapour diffusion

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221 SiO2-Ag+Chlorex vs SilverSulfaDiazine: An 'in vitro' and 'in vivo' Silver Challenge

Authors: Roberto Cassino, Valeria Dissette, Carlo Alberto Bignozzi, Daniele Pazzi

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Background and Aims: The aim of this work was to investigate, both ‘in vitro’ and ‘in vivo’, if the new SCX technology (SiO2-Ag+Chlorex) can easily defeat infections and it is really more effective than SSD (SilverSulfaDiazine). ‘In vitro’ methods: we tested ‘in vitro’ the effectiveness of both silver materials using a pool of 5 strains: Pseudomonas Aeruginosa, Staphylococcus aureus, Escherichia Coli, Enterococcus hirae and Candida Albicans. 100 µl of this pool have been seeded on Petri dishes and kept for 24 hours in incubation at 37 C°. ‘In vivo’ methods: we enrolled patients with multiple infectious chronic wounds (according with cutting & harding criteria for infection); after a qualitative evaluation of the wounds bacterial population, taking a sample by plug, we included in the study 6 patients for a total of 10 wounds, infected by one or more of the microorganisms used for the ‘in vitro’ test. The protocol consisted of a treatment with a spray powder of SSD every 48 hours for 14 days; in case of worsening we should have to start a new treatment with a spray powder containing silicon dioxide, ionic silver and chlorexidine (SiO2-Ag+Chlorex) every 48 hours for 14 days. We evaluated the number of clinical signs of infection and the disappearance or not of the wound edge erithema. ‘In vitro’ results: SSD demonstrated a wide zone of inhibition within 24 hours, but after 5 days there was no more signs of inhibition; on the contrary SCX had a good inhibition ring that lasted more than 5 days. ‘In vivo’ results: all wounds treated with SSD got worse; the signs of infection increased and the wound edge erithema did not disappear. According with the protocol, we treated then all wounds with SCX and they all improved within the period of observation with complete disappearance of clinical signs of infection and no more wound edge erithema. Conclusions: the study demonstrated the effectiveness of SiO2-Ag+Chlorex, especially in terms of long lasting antimicrobial action. We had the same results ‘in vitro’, so that there has been a perfect correspondence between the laboratory outcomes and the clinical ones.

Keywords: chronic wounds, infections, ionic silver, SSD

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220 Assessment of Spatial and Temporal Variations of Some Biological Water Quality Parameters in Mat River, Albania

Authors: Etleva Hamzaraj, Eva Kica, Anila Paparisto, Pranvera Lazo

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Worldwide demographic developments of recent decades have been associated with negative environmental consequences. For this reason, there is a growing interest in assessing the state of natural ecosystems or assessing human impact on them. In this respect, this study aims to evaluate the change in water quality of the Mat River for a period of about ten years to highlight human impact. In one year, period of study, several biological and environmental parameters are determined to evaluate river water quality, and the data collected are compared with those of a similar study in 2007. Samples are collected every month in five stations evenly distributed along the river. Total coliform bacteria, the number of heterotrophic bacteria in water, and benthic macroinvertebrates are used as biological parameters of water quality. The most probable number index is used for evaluation of total coliform bacteria in water, while the number of heterotrophic bacteria is determined by counting colonies on plates with Plate Count Agar, cultivated with 0.1 ml sample after series dilutions. Benthic macroinvertebrates are analyzed by the number of individuals per taxa, the value of biotic index, EPT Richness Index value and tolerance value. Environmental parameters like pH, temperature, and electrical conductivity are measured onsite. As expected, the bacterial load was higher near urban areas, and the pollution increased with the course of the river. The maximum concentration of fecal coliforms was 1100 MPN/100 ml in summer and near the most urbanized area of the river. The data collected during this study show that after about ten years, there is a change in water quality of Mat River. According to a similar study carried out in 2007, the water of Mat River was of ‘excellent’ quality. But, according to this study, the water was classified as of ‘excellent’ quality only in one sampling site, near river source, while in all other stations was of ‘good’ quality. This result is based on biological and environmental parameters measured. The human impact on the quality of water of Mat River is more than evident.

Keywords: water quality, coliform bacteria, MPN index, benthic macroinvertebrates, biotic index

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219 Comparative Study of Antimicrobial, Antioxidant and Physicochemical Properties of Four Culinary Herbs Grown in Sri Lanka

Authors: Thilini Kananke

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Culinary herbs have long been considered as significant dietary sources of many potential health-promoting compounds. The present research focused on analysis of antimicrobial, antioxidant and physicochemical properties in selected four culinary herbs namely Murraya koenigii (Curry leaves), Pandanus amaryllifolius (Pandan leaves), Cymbopogon citrates (Lemon grass leaves), and Mentha Piperita (Minchi leaves) obtained from several market sites in Ratnapura District, Sri Lanka. The antimicrobial activity of ethanolic, chloroform and distilled water extracts of culinary herbs were evaluated against the strains of Staphylococcus aureus, Salmonella typhi and Shigella spp. Total phenolic content and the radical scavenging activity (using DPPH assay) of culinary herbs were determined. Four heavy metals (Cu, Cd, Pb and Fe) were analyzed in the selected culinary herbs using the atomic absorption spectroscopy (AAS). Proximate compositions of the selected herbs were analyzed using AOAC official methods. Antimicrobial activity of all selected culinary herbs showed relativity high inhibition zones against S. aureus. Pandan leaves showed the least antimicrobial activity against selected bacterial strains compared with other culinary herbs. Both the highest radical scavenging activity (lower IC50 value) and the total phenolic content (25.57 ±3.54µg GAE/100g) were reported in Mentha piperita extract. The highest concentrations of Cu, Fe and Cd were reported in Curry leaves (29.15 mg/kg), Lemon grass leaves (257.98 mg/kg) and Pandan leaves (6.05 mg/kg) respectively. The heavy metal contents detected in all culinary herbs were below the permitted limits set by WHO/FAO, except Cd. The highest moisture (85.00±0.00%) and fiber (10.66± 2.00%) contents were found in Pandan leaves, while the highest protein (8.94±0.29%), fat (12.3± 2.52%) and ash (3.50± 0.17%) contents were reported in curry leaves. The information obtained from this study highlights the importance of further investigation of other antioxidant, antimicrobial and health promoting compounds of culinary herbs available in Sri Lanka for a detailed comparison.

Keywords: antimicrobial, antioxidant, culinary herbs, proximate analysis

Procedia PDF Downloads 179
218 Potential of Native Microorganisms in Tagus Estuary

Authors: Ana C. Sousa, Beatriz C. Santos, Fátima N. Serralha

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The Tagus estuary is heavily affected by industrial and urban activities, making bioremediation studies crucial for environmental preservation. Fuel contamination in the area can arise from various anthropogenic sources, such as oil spills from shipping, fuel storage and transfer operations, and industrial discharges. These pollutants can cause severe harm to the ecosystem and the organisms, including humans, that inhabit it. Nonetheless, there are always natural organisms with the ability to resist these pollutants and transform them into non-toxic or harmless substances, which defines the process of bioremediation. Exploring the microbial communities existing in soil and their capacity to break down hydrocarbons has the potential to enhance the development of more efficient bioremediation approaches. The aim of this investigation was to explore the existence of hydrocarbonoclastic microorganisms in six locations within the Tagus estuary, three on the north bank: Trancão River, Praia Fluvial do Cais das Colinas and Praia de Algés, and three on the south bank: Praia Fluvial de Alcochete, Praia Fluvial de Alburrica, and Praia da Trafaria. In all studied locations, native microorganisms of the genus Pseudomonas were identified. The bioremediation rate of common hydrocarbons like gasoline, hexane, and toluene was assessed using the redox indicator 2,6-dichlorophenolindophenol (DCPIP). Effective hydrocarbon-degrading bacterial strains were identified in all analyzed areas, despite adverse environmental conditions. The highest bioremediation rates were achieved for gasoline (68%) in Alburrica, hexane (65%) in Algés, and toluene (79%) in Algés. Generally, the bacteria demonstrated efficient degradation of hydrocarbons added to the culture medium, with higher rates of aerobic biodegradation of hydrocarbons observed. These findings underscore the necessity for further in situ studies to better comprehend the relationship between native microbial communities and the potential for pollutant degradation in soil.

Keywords: biodegradability rate, hydrocarbonoclastic microorganisms, soil bioremediation, tagus estuary

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217 Correlation Between HIV/AIDS Stage With Oral Health, Dentition, and Periodontal Status

Authors: Eriselda Simoni, Leonard Simoni, Endri Paparisto, Laureta Flaga, Silvana Bara, Edit Xhajanka, Arjan Harxhi

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Background: Some pathologies are encountered more often in HIV/ AIDS, such as those with bacterial, fungal, viral, and neoplastic causes, but what has been more noticeable in recent years is the increased and more aggressive manifestation of periodontal disease and oral caries. Our purpose is to investigate the correlation between the HIV/AIDS stage and CD4 level with oral health, dentition, and periodontal status. Materials and Methods: We conducted a prospective observational study that included 35 patients newly diagnosed with HIV/AIDS and underwent an oral examination at the University Dental Clinic in Tirana, Albania, in the period April - July 2024. This study evaluated the basic demographic, laboratory characteristics, oral hygiene, and the presence of oral lesions. The dentition status was assessed with the values DT (decay teeth), FT (filled teeth), and MT (missing teeth) presented as DMFT. The periodontal status was evaluated through a periodontal probe measuring CPI (community periodontal index) and LOA (loss of attachment) as recommended by the WHO Oral Health Assessment Form 2013. The Pearson Correlation Coefficient (r) was used to evaluate the relationship between levels of CD4+ and DMF, CD4+ and CPI, and CD4+ and LOA. The P value ≤ 0.05 was considered statistically significant. Results: 80% of patients included were males with a mean age of 35.8 years. 8.6% of patients were categorized as HIV stage I, 28.6% as stage II, and 62.8% as HIV stage III/AIDS. The mean level value of CD4+ was 266.2 cells/mm3 and the rapport CD4+/ WBC (White Blood Cells) was 15.7%. Most patients (57.2%) used toothbrushes less than 1 time a day. An important negative correlation was found between CD4+ and dentition and periodontal status. A lower level of CD4+ was correlated with a higher DMFT, CPI, and LOA, respectively coefficient (r) for CD4/DMFT = -0.52, p =0.01, (r) for CD4/CPI= - 0.38, p=0.024 and (r) for CD4/LOA= - 0.37, p=0.029. Conclusions: In our study, it was documented that patients with HIV/AIDS had worse oral health, an important negative correlation between CD4+ and dentition and periodontal status. A lower level of CD4+ was correlated with a worse dentition status (higher DMFT), and poor periodontal health (higher CPI and LOA). The monitoring and treatment of oral pathologies can be important in early HIV/AIDS diagnoses and treatment.

Keywords: HIV/AIDS, oral health, dentition, periodontal

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216 Culturable Microbial Diversity and Adaptation Strategy in the Jutulsessen and Ahlmannryggen of Western Dronning Maud Land, Antarctica

Authors: Shiv Mohan Singh, Gwyneth Matcher

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To understand the culturable microbial composition and diversity patterns, soil samples were collected from inland nunataks of Jutulsessen and Ahlmannryggen ranges in Dronning Maud Land, Antarctica. 16S rRNA, ITS and the D1/D2 domain sequencing techniques were used for characterization of microbial communities of these geographical areas. The total 37 species of bacteria such as Arthrobacter agilis, Acinetobacter baumannii, Arthrobacter flavus, Arthrobacter ginsengisoli, Arthrobacter oxydans, Arthrobacter oryzae, Arthrobacter polychromogenes, Arthrobacter sulfonivorans, Bacillus altitudinis, Bacillus cereus, Bacillus paramycoides, Brevundimonas vesicularis, Brachybacterium rhamnosum, Curtobacterium luteum, Dermacoccus nishinomiyaensis, Dietzia aerolata, Janibacter indicus, Knoellia subterranean, Kocuria palustris, Kytococcus aerolatus, Lysinibacillus sphaericus, Microbacterium phyllosphaerae, Micrococcus yunnanensis, Methylobacterium rhodesianum, Moraxella osloensis, Paracoccus acridae, Pontibacter amylolyticus, Pseudomonas hunanensis, Pseudarthrobacter siccitolerans, Pseudarthrobacter phenanthrenivorans, Rhodococcus aerolatus, Rhodococcus sovatensis, Sphingomonas daechungensis, Sphingomonas sanguinis, Stenotrophomonas pavanii, Staphylococcus gallinarum, Staphylococcus arlettae and 9 species of fungi such as Candida davisiana, Cosmospora arxii, Geomyces destructans, Lecanicillium muscarium, Memnoniella humicola, Paecilomyces lilacinus, Pseudogymnoascus verrucosus, Phaeophlebiopsis ignerii and Thyronectria caraganae were recorded. Fatty acid methyl esters (FAME) analyses of representative species of each genus have shown predominance branched and unsaturated fatty acids indicate its adaptation strategy in Antarctic cold environment. To the best of our knowledge, this is the first record of culturable bacterial communities from Jutulsessen and Ahlmannryggen ranges in Western Dronning Maud Land, Antarctica.

Keywords: antarctica, microbe, adaptation, polar

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215 Prevalence and Antibiotic Resistance of Bacteria Isolated from Farmers’ Market Fruits and Vegetables Collected from Frostburg and Cumberland Areas in Maryland

Authors: Kumudini Apsara Munasinghe, Devin Gregory Lissau, Ryan Thomas Wade

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Fresh fruits and vegetables are rich in vitamins, minerals, and fibers and help maintain a healthy weight over high-calorie food. Eating fruits and vegetables protects us from free radicals produced by metabolic reactions and safeguards us from cardiovascular disease and cancer. However, there has been an increased concern about foodborne diseases tied to contaminated farmers’ market produce. In addition, very little information is available about the contribution of eating raw fruits and vegetables to human exposure to antibiotic-resistant bacteria. This research aims to identify bacteria isolated from farmers’ market fruits and vegetables and understand their antibiotic resistance. Vegetables and fruits were collected from farmers’ markets around Frostburg and Cumberland areas in Maryland and transported to the microbiology lab at Frostburg State University for the isolation of bacteria. Bacteria were extracted from tomatoes, cucumber, strawberry, and lettuce using Tryptic soy broth overnight at 37°C, and Tryptic Soy agar was used for the streak plate technique to isolate bacteria. Pure cultures were used to identify bacteria using biochemical reactions after conducting Gram staining technique. The research used many biochemical reactions, including Mannitol Salt agar, MacConkey agar, and Eosin Methylene blue agar, for identification. Antibiotic sensitivity was tested for many different types of antibiotics, including amoxicillin, penicillin, tetracycline, ampicillin, and erythromycin. Most prevalent bacteria in the isolates were Staphylococcus, Bacillus, Micrococcus, Enterococcus, Enterobacter, Citrobacter, and other bacteria from the family Enterobacteriaceae. The data obtained from this research will be useful to educate and train farmers and individuals involved in post-harvest processes such as transportation and selling in farmers’ markets. Further results for bacterial antibiotic resistance will be obtained, and unculturable bacteria will be identified by next-generation DNA sequencing.

Keywords: antibiotic resistance, farmers markets, fruits, bacteria, vegetables

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214 Prevalence and Drug Susceptibility Profiles of Bacterial Urinary Tract Infections Isolated among Diabetes Mellitus Patients at Bosaso Health Centers

Authors: Said Abdirasak Abidrahman, Ibrahim Mohamed

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Background: Urinary Tract Infections (UTIs) are the commonest infections described among diabetes mellitus patients. More often, empirical antimicrobial therapy is initiated before the laboratory results are made available with minimal treatment success. The knowledge of the etiology and antibiotic susceptibility patterns of the organisms causing urinary tract infections among diabetes mellitus patients remains scarce, despite its vitality. This study sought to determine the prevalence, bacteria species, and drug susceptibility patterns of common causes of urinary tract infections among diabetes mellitus patients attending Bosaso health centers. Materials and methods: We conducted a cross-sectional study involving adult diabetic patients at Bosaso health centers between the months of May and July 2020. Laboratory assay of mid-stream urine samples was done to isolate bacteria causes of UTIs. These were biochemically identified using Gram stain, Kligler iron agar (KIA), Indole test, citrate, urea, coagulase, catalase, motility agar, and lysine iron agar. Their antibiotic susceptibility pattern for the isolated organisms was made for Ampicillin 10μg, Ciprofloxacin 5μg, Cotrimoxazole 25μg, Gentamycin 10μg, Ceftriaxone 10μg, and determined using the Kirby Bauer Disc Diffusion method. Results: Of 177 participants, 69 (39.0%) were males and 108 (61.0%) were females. Their mean age was 33.1 years (range; 18-67 years). Of these, 14.7% (26/177) of the samples revealed significant growth (>= 105 CFU/mL) giving a prevalence of 14.9 % (95% CI: 10.6 to 16.3). The organisms isolated were Escherichia coli -50% (N=13), Klebsiella pneumonia 30.8% (N=8), Staphylococcus aureus 15.4% (N=4), and unidentified organism 3.8% (N=1), and these were associated with such socio-demographic factors like history of catheterization and sexual activity. Antibiotic susceptibility to the commonly used agents for treating UTIs indicated higher sensitivity to Gentamicin and Ceftriaxone.

Keywords: antimicrobials, bacteria, urinary tract infections, diabetes

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213 Staphylococcal Enterotoxins Play an Important Role in Clinical Signs in Bovine Mastitis

Authors: Stéfani T. A. Dantas, Laura T. S. Takume, Bruna F. Rossi, Érika R. Bonsaglia, Ivana G. Castilho, José C. F. Pantoja, Ary Fernandes Júnior, Juliano L. Gonçalves, Marcos V. Santos, Rinaldo A. Mota, Vera L. M. Rall

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Staphylococcus aureus is one of the main pathogens causing contagious bovine mastitis, being more frequently isolated from subclinical form, although the clinical form also occurs. Clinical mastitis cause visual signs, such as swelling, fever, hardening of the mammary gland, or any change in the characteristics of the milk. Considering the subclinical type, there are no visible signs in the animal nor changes in the milk. S. aureus has many important virulence factors for the establishment of its pathogenicity in animals, such as enterotoxins, which are also responsible for foodborne poisoning. Our objective is to perform a comparative analysis between 103 isolates of S. aureus, obtained from the milk of cows with clinical mastitis and 103 more, from subclinical type, in relation to the presence of these enterotoxins and verify if their presence plays an important role in the signs of illness. We will investigate all enterotoxins described till now, such as sea-see, seg-sez, sel26, sel 27, se01, and se02 (This study was approved by the Sao Paulo State University Animal Use Ethics Committee, No. 0136/2017). For the PCR assay, we used Illustra Bacteria Mini Spin Kit for bacterial DNA. At this moment, we have already tested sea-see, seg-ser, sew, and sex, and the results have already been submitted to Fisher Exact Probability Test or Chi-square Test. Considering the isolates obtained from clinical mastitis, the most frequent enterotoxins were selw (99%), selx (78%) and selh (50.5%), and sec, see, sej, sell, selp,and ser were absent. Among the subclinics, selw (82.5%) selm (15.5%) and selx (14.6%) were the most frequent, and sea-see, seg, sei-sel, sem-ser were absent. We have already observed statistically significant differences for seb, seg, seh, sei, selo, selu, selw and selx. Other interesting results were the low number of genes in each isolate from subclinical mastitis [0 genes: 14 (13.6%); 1 gene: 55 (53.4%); 2 genes: 33 (32%) or 3: 1 (0.97%)] compared to clinical isolates [1 gene: 5 (4.9%); 2 genes: 29 (28.1%); 3 genes: 38 (36.9%); 4 genes: 14 (13.6%); 5 genes: 5 (4.9%); 6 genes: 4 (3.9%); 7 genes: 5 (4.9%); 8 genes: 2 (1.9%) and 9 genes: 1 (1%)]. Based on these results, we can conclude that enterotoxins indeed play an important role in clinical signs in cattle with mastitis.

Keywords: mastitis, S. aureus, PCR, staphylococcal enterotoxin

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212 Physicochemical and Functional significance of Two Lychee (Litchi chinensis Sonn.) Cultivars Gola and Surakhi from Pakistan

Authors: Naila Safdar, Faria Riasat, Azra Yasmin

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Lychee is an emerging fruit crop in Pakistan. Two famous cultivars of lychee, Gola and Surakhi, were collected from Khanpur Orchard, Pakistan and their whole fruit (including peel, pulp and seed) was investigated for pomological features and therapeutic activities. Both cultivars differ in shape and size with Gola having large size (3.27cm length, 2.36cm width) and more flesh to seed ratio (8.65g). FTIR spectroscopy and phytochemical tests confirmed presence of different bioactive compounds like phenol, flavonoids, quinones, anthraquinones, tannins, glycosides, and alkaloids, in both lychee fruits. Atomic absorption spectroscopy indicated an increased amount of potassium, magnesium, sodium, iron, and calcium in Gola and Surakhi fruits. Small amount of trace metals, zinc and copper, were also detected in lychee fruit, while heavy metals lead, mercury, and nickel were absent. These two lychee cultivars were also screened for antitumor activity by Potato disc assay with maximum antitumor activity shown by aqueous extract of Surakhi seed (77%) followed by aqueous extract of Gola pulp (74%). Antimicrobial activity of fruit parts was checked by agar well diffusion method against six bacterial strains Enterococcus faecium, Enterococcus faecalis, Staphylococcus aureus, Bacillus subtilis, Bacillus sp. MB083, and Bacillus sp. MB141. Highest antimicrobial activity was shown by methanolic extract of Gola pulp (27mm ± 0.70) and seed (19.5mm ± 0.712) against Enterococcus faecalis. DPPH scavenging assay revealed highest antioxidant activity by aqueous extract of Gola peel (98.10%) followed by n-hexane extract of Surakhi peel (97.73%). Results obtained by reducing power assay also corroborated with the results of DPPH scavenging activity.

Keywords: antimicrobial evaluation, antitumor assay, gola, phytoconstituents, reactive oxygen species, Surakhi

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211 Field Study of Chlorinated Aliphatic Hydrocarbons Degradation in Contaminated Groundwater via Micron Zero-Valent Iron Coupled with Biostimulation

Authors: Naijin Wu, Peizhong Li, Haijian Wang, Wenxia Wei, Yun Song

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Chlorinated aliphatic hydrocarbons (CAHs) pollution poses a severe threat to human health and is persistent in groundwater. Although chemical reduction or bioremediation is effective, it is still hard to achieve their complete and rapid dechlorination. Recently, the combination of zero-valent iron and biostimulation has been considered to be one of the most promising strategies, but field studies of this technology are scarce. In a typical site contaminated by various types of CAHs, basic physicochemical parameters of groundwater, CAHs and their product concentrations, and microbial abundance and diversity were monitored after a remediation slurry containing both micron zero-valent iron (mZVI) and biostimulation components were directly injected into the aquifer. Results showed that groundwater could form and keep low oxidation-reduction potential (ORP), a neutral pH, and anoxic conditions after different degrees of fluctuations, which was benefit for the reductive dechlorination of CAHs. The injection also caused an obvious increase in the total organic carbon (TOC) concentration and sulfate reduction. After 253 days post-injection, the mean concentration of total chlorinated ethylene (CEE) from two monitoring wells decreased from 304 μg/L to 8 μg/L, and total chlorinated ethane (CEA) decreased from 548 μg/L to 108 μg/L. Occurrence of chloroethane (CA) suggested that hydrogenolysis dechlorination was one of the main degradation pathways for CEA, and also hints that biological dechlorination was activated. A significant increase of ethylene at day 67 post-injection indicated that dechlorination was complete. Additionally, the total bacterial counts increased by 2-3 orders of magnitude after 253 days post-injection. And the microbial species richness decreased and gradually changed to anaerobic/fermentative bacteria. The relative abundance of potential degradation bacteria increased corresponding to the degradation of CAHs. This work demonstrates that mZVI and biostimulation can be combined to achieve the efficient removal of various CAHs from contaminated groundwater sources.

Keywords: chlorinated aliphatic hydrocarbons, groundwater, field study, zero-valent iron, biostimulation

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210 Impure Water, a Future Disaster: A Case Study of Lahore Ground Water Quality with GIS Techniques

Authors: Rana Waqar Aslam, Urooj Saeed, Hammad Mehmood, Hameed Ullah, Imtiaz Younas

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This research has been conducted to assess the water quality in and around Lahore Metropolitan area on the basis of three different land uses, i.e. residential, commercial, and industrial land uses. For this, 29 sample sites have been selected on the basis of simple random sampling technique. Samples were collected at the source (WASA tube wells). The criteria for selecting sample sites are to have a maximum concentration of population in the selected land uses. The results showed that in the residential land use the proportion of nitrate and turbidity is at their highest level in the areas of Allama Iqbal Town and Samanabad Town. Commercial land use of Gulberg and Data Gunj Bakhsh Town have highest level of proportion of chlorides, calcium, TDS, pH, Mg, total hardness, arsenic and alkalinity. Whereas in industrial type of land use in Ravi and Wahga Town have the proportion of arsenic, Mg, nitrate, pH, and turbidity are at their highest level. The high rate of concentration of these parameters in these areas is basically due to the old and fractured pipelines that allow bacterial as well as physiochemical contaminants to contaminate the portable water at the sources. Furthermore, it is seen in most areas that waste water from domestic, industrial, as well as municipal sources may get easy discharge into open spaces and water bodies, like, cannels, rivers, lakes that seeps and become a part of ground water. In addition, huge dumps located in Lahore are becoming the cause of ground water contamination as when the rain falls, the water gets seep into the ground and impures the ground water quality. On the basis of the derived results with the help of Geo-spatial technology ACRGIS 9.3 Interpolation (IDW), it is recommended that water filtration plants must be installed with specific parameter control. A separate team for proper inspection has to be made for water quality check at the source. Old water pipelines must be replaced with the new pipelines, and safe water depth must be ensured at the source end.

Keywords: GIS, remote sensing, pH, nitrate, disaster, IDW

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209 Non Chemical-Based Natural Products in the Treatment and Control of Disease in Fish

Authors: Albert P. Ekanem, Austin I. Obiekezie, Elizabeth X. Ntia

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Introduction: Some African plants and bile from animals have shown efficacies in the treatment and control of diseases in farmed fish. The background of the study is based on the fact the African rain forest is blessed with the abundance of medicinal plants that should be investigated for their use in the treatment of diseases. The significance of the study is informed by the fact that chemical-based substances accumulate in the tissues of food fish, thereby reducing the food values of such products and moreover, the continuous use of chemotherapeutics in the aquatic environments tends to degrade the affected environment. Methodology: Plants and animal products were extracted, purified and applied under in vitro and in vivo conditions to the affected organisms. Effective plants and bills were analyzed for biologically active substances responsible for the activities by both qualitative and HPLC methods. Results: Extracts of Carica papaya and Mucuna pruriens were effective in the treatment of Ichthyophthiriasis in goldfish (Carassius auratus auratus) with high host tolerance. Similarly, ectoparasitic monogeneans were effectively dislodged from the gills and skin of goldfish by the application of extracts of Piper guineense at therapeutic concentrations. Artemesia annua with known antimalarial activities in human was also effective against fish monogenean parasites of Clarias gariepinus in a concentration-related manner without detriments to the host. Effective antibacterial activities against Aeromonas and Pseudomonas diseases of the African catfish (Heterobranchus longifilis) were demonstrated in some plants such as Phylanthus amarus, Allium sativum, A. annua, and Citrus lemon. Bile from some animals (fish, goat, chicken, cow, and pig) showed great antibacterial activities against some gastrointestinal bacterial pathogens of fish. Conclusions: African plants and some animal bile have shown potential promise in the treatment of diseases in fish and other aquatic animals. The use of chemical-based substances for control of diseases in the aquatic environments should be restricted.

Keywords: control, diseases, fish, treatment

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208 Degradation of Commercial Polychlorinated Biphenyl Mixture by Naturally Occurring Facultative Microorganisms via Anaerobic Dechlorination and Aerobic Oxidation

Authors: P. M. G. Pathiraja, P. Egodawatta, A. Goonetilleke, V. S. J. Te'o

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The production and use of Polychlorinated biphenyls (PCBs), a group of synthetic halogenated hydrocarbons have been restricted worldwide due to its toxicity and categorized as one of the twelve priority persistent organic pollutants (POP) by the Stockholm Convention. Low reactivity and high chemical stability of PCBs have made them highly persistent in the environment and bio-concentration and bio-magnification along the food chain contribute to multiple health impacts in humans and animals. Remediating environments contaminated with PCBs is a challenging task for decades. Use of microorganisms for remediation of PCB contaminated soils and sediments have been widely investigated due to the potential of breakdown these complex contaminants with minimum environmental impacts. To achieve an effective bioremediation of polychlorinated biphenyls (PCBs) contaminated environments, microbes were sourced from environmental samples and tested for their ability to hydrolyze PCBs under different conditions. Comparison of PCB degradation efficiencies of four naturally occurring facultative bacterial cultures isolated through selective enrichment under aerobic and anaerobic conditions were simultaneously investigated in minimal salt medium using 50 mg/L Aroclor 1260, a commonly used commercial PCB mixture as the sole source of carbon. The results of a six-week study demonstrated that all the tested facultative Achromobacter, Ochrobactrum, Lysinibacillus and Pseudomonas strains are capable of degrading PCBs under both anaerobic and aerobic conditions while assisting hydrophobic PCBs to make solubilize in the aqueous minimal medium. Overall, the results suggest that some facultative bacteria are capable of effective in degrading PCBs under anaerobic conditions through reductive dechlorination and under aerobic conditions through oxidation. Therefore, use of suitable facultative microorganisms under combined anaerobic-aerobic conditions and combination of such strains capable of solubilization and breakdown of PCBs has high potential in achieving higher PCB removal rates.

Keywords: bioremediation, combined anaerobic-aerobic degradation, facultative microorganisms, polychlorinated biphenyls

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207 Extracellular Production of the Oncolytic Enzyme, Glutaminase Free L-Asparaginase, from Newly Isolated Streptomyces Olivaceus NEAE-119: Optimization of Culture Conditions Using Response Surface Methodology

Authors: Noura El-Ahmady El-Naggar

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Among the antitumour drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product(1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen nutritional variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4.7H2O, NaCl and FeSO4. 7H2O) were screened using Plackett–Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age and agitation speed) were further optimized by the central composite face-centered design -response surface methodology. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase in the culture filtrate of Streptomyces olivaceus NEAE-119: Dextrose 3g, starch 20g, L-asparagine 10g, KNO3 1g, K2HPO4 1g, MgSO4.7H2O 0.1g, NaCl 0.1g, pH 7, temperature 37°C, agitation speed 200 rpm/min, inoculum size 4%, v/v, inoculum age 72 h and fermentation period 5 days.

Keywords: Streptomyces olivaceus NEAE-119, glutaminase free L-asparaginase, production, Plackett-Burman design, central composite face-centered design, 16S rRNA, scanning electron microscope

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